Darribere T, Boucaut J C, Keil-Dlouha V
Biochim Biophys Acta. 1982 Apr 7;686(2):145-52. doi: 10.1016/0005-2736(82)90106-7.
Surface proteins in the first embryonic stages (8-32 cells, morula, blastula, early and late gastrula) of Pleurodeles waltlii were selectively labelled by 125I using lactoperoxidase and glucose/glucose oxidase. Iodination was effected either on non-dissociated embryos or after their dissociation with EDTA. On the outer surface of non-dissociated embryos the two-dimensional electrophoresis revealed only three groups of 125I-labelled proteins which did not change during all studied stages. Quite different results were obtained with the cells of dissociated embryos. In addition to the iodinated proteins of the embryonic outer surface seven major iodinated proteins were identified. These proteins originate from the regions of cell-cell contacts in intact embryo. Their two-dimensional pattern in dissociated cells changes between stages 8-32 cells and morula. The next important difference was observed during gastrulation, which corresponds in Pleurodeles waltlii to the first morphogenetic movements. Therefore the outside and inside cell surfaces of embryo are different already at stage 8-32 cells (and probably earlier), before the first step of morphogenesis. The changes of cell surface proteins at early embryonal development take place inside the embryo, in the regions of cell-cell interactions.
使用乳过氧化物酶和葡萄糖/葡萄糖氧化酶,用¹²⁵I对滑蹠螈胚胎发育早期阶段(8 - 32细胞期、桑椹胚、囊胚、原肠胚早期和晚期)的表面蛋白进行选择性标记。碘化反应在未解离的胚胎上进行,或在胚胎用乙二胺四乙酸(EDTA)解离后进行。在未解离胚胎的外表面,二维电泳仅显示三组¹²⁵I标记的蛋白,在所有研究阶段均未发生变化。解离胚胎的细胞得到了截然不同的结果。除了胚胎外表面的碘化蛋白外,还鉴定出七种主要的碘化蛋白。这些蛋白源自完整胚胎中细胞 - 细胞接触区域。它们在解离细胞中的二维图谱在8 - 32细胞期和桑椹胚阶段之间发生变化。在下胚层形成过程中观察到了下一个重要差异,这在滑蹠螈中对应于第一次形态发生运动。因此,在形态发生的第一步之前,胚胎的外部和内部细胞表面在8 - 32细胞期(可能更早)就已经不同。胚胎早期发育过程中细胞表面蛋白的变化发生在胚胎内部,即细胞 - 细胞相互作用的区域。